WO2017037829A1

WO2017037829A1 - Method for quantitative analysis of high-molecular-weight compound, and data processing device for said quantitative analysis

Info

Publication number: WO2017037829A1
Application number: PCT/JP2015/074652
Authority: WO
Inventors: 修一中家; 慎治船津
Original assignee: 株式会社島津製作所
Priority date: 2015-08-31
Filing date: 2015-08-31
Publication date: 2017-03-09
Also published as: US20180337028A1; EP3364184A4; US10438785B2; JPWO2017037829A1; JP6508342B2; CN107923873B; CN107923873A; EP3364184A1

Abstract

When an analyst inputs the structure of each of sugar chains supposed to be contained in a sample, a supposed ionic valence, a common product ion m/z, etc., then a precursor m/z calculation unit (301) calculates the m/z of a precursor ion derived from the sugar chain and a method file creation unit (305) creates a method file including MRM transitions. A polyvalent-ion information file creation unit (303) creates a file which links a unique ID of the sugar-chain structure to the precursor ion m/z and the product ion m/z. After completion of a measurement, a chromatogram creation unit (34) makes a mass chromatogram for each MRM transition, and a peak-area addition unit (311) adds up the areas of peaks of the mass chromatograms for the MRM transitions corresponding to the same sugar chain by reference to the linking file. A quantitative-value calculation unit (312) determines the proportion of each sugar chain present, as a quantitative value on the basis of the total value. A quantitative output information creation unit (39) converts the quantitative values into a graph, and the graph is output to a display unit (5). Thus, simultaneous multi-component analysis of a compound which is apt to generate a polyvalent ion, such as a sugar chain, can be made with LC-MS.

Description

Method for quantitative analysis of polymer compound and data processing apparatus for the quantitative analysis

The present invention relates to a quantitative analysis method for quantifying a high molecular compound such as a sugar chain or a glycopeptide in a sample and a data processing apparatus for the quantitative analysis, and more specifically, a mass spectrometer capable of MS / MS analysis. The present invention relates to a method for quantifying a polymer compound in a sample and a data processing apparatus therefor.

For quantitative analysis of low molecular weight compounds such as pharmaceuticals and agricultural chemicals, multiple reaction monitoring (MRM = Multiple Reaction) using a liquid chromatograph mass spectrometer (LC-MS) that combines a liquid chromatograph and a tandem quadrupole mass spectrometer Monitoring) is often used. In MRM measurement, the influence of contaminant components can be eliminated by a two-stage mass separator (quadrupole mass filter), so there are contaminant components with retention times close to those of the target component. Even if it cannot be separated, the quantitative analysis of the target component can be performed with high accuracy. In general, in MRM measurement by LC-MS, a plurality of combinations of precursor ions and product ions (generally referred to as MRM transitions) can be set in one measurement time range. Dozens of components can be quantified. Such an analysis method is called multi-component simultaneous analysis, and has been widely used in recent years for inspection of residual agricultural chemicals and inspection of pollutants in environmental water.

On the other hand, liquid chromatographs that still use ultraviolet absorption detectors or fluorescence detectors as detectors are still frequently used for quantitative analysis of polymer compounds such as sugar chains and glycopeptides (see Non-Patent Documents 1 and 2). ). One of the reasons is that, in an ion source such as an electrospray ionization (ESI) method generally used in LC-MS, the valence is not 1 when ionizing a polymer compound such as a sugar chain or a glycopeptide. This is because a plurality of types of valence ions are likely to be generated, so that it is not a simple combination of one precursor ion and one product ion for each component like a low molecular compound. Furthermore, as is well known, there is a problem in that the sugar chain structure has non-uniformity, and the ionic valence detected by the sample varies greatly. For this reason, unlike the quantitative analysis of low molecular weight compounds, multi-component simultaneous analysis using LC-MS is difficult for quantitative analysis of high molecular weight compounds such as sugar chains and glycopeptides. As a result, it is necessary to perform a quantitative analysis for each compound using a liquid chromatograph with an ultraviolet absorption detector or a fluorescence detector as the detector, resulting in low analysis throughput, reduced analysis time, and labor. Reduction has become a major issue.

JP 2014-66704 A

The present invention has been made in order to solve the above-mentioned problems, and its object is to provide a mass spectrometer capable of performing MS / MS analysis such as a liquid chromatograph and a tandem quadrupole mass spectrometer, for example. Can improve the throughput of quantitative analysis of high-molecular compounds by enabling multi-component simultaneous analysis of high-molecular compounds such as sugar chains and glycopeptides by using MRM measurement in a device combined with It is to provide a method for quantitative analysis of a polymer compound that can be produced and a data processing device for carrying out the method.

The quantitative analysis method of a polymer compound according to the present invention made to solve the above problems is a quantitative analysis method for quantifying a polymer compound in a sample using a mass spectrometer capable of MS / MS analysis. There,
a) With respect to a polymer compound to be analyzed or assumed to be analyzed, a plurality of precursor ions having different ionic valences derived from the compound and a plurality of MRM measurements targeting one common product ion are used. A peak information acquisition step for calculating a total value of the peak areas appearing in the obtained plurality of mass chromatograms,
b) Using the total value obtained in the peak information acquisition step, a quantitative step for performing a quantitative calculation of a polymer compound that is an analysis target or assumed as an analysis target;
It is characterized by having.

In addition, a data processing apparatus for quantitative analysis of a polymer compound according to the present invention, which has been made to solve the above problems, is an apparatus for carrying out the above quantitative analysis method, and is capable of performing MS / MS analysis. A data processing device for quantifying a polymer compound in a sample based on data obtained using an analysis device,
a) With respect to a polymer compound to be analyzed or assumed to be analyzed, a plurality of precursor ions having different ionic valences derived from the compound and a plurality of MRM measurements targeting one common product ion are used. A peak information acquisition unit for calculating a total value of the peak areas appearing in the obtained plurality of mass chromatograms;
b) Using the total value obtained by the peak information acquisition unit, a quantitative processing unit that performs a quantitative calculation of a polymer compound that is an analysis target or is assumed to be an analysis target;
It is characterized by having.

Typically, as a mass spectrometer for acquiring data to be processed in the polymer compound quantitative analysis method and the data processing apparatus according to the present invention, a quadrupole mass filter is provided before and after the collision cell. Arranged tandem quadrupole mass spectrometer, Q-TOF mass spectrometer in which the subsequent quadrupole mass filter is replaced with a time-of-flight mass spectrometer in the tandem quadrupole mass spectrometer, or a collision cell And a TOF / TOF mass spectrometer in which two stages of time-of-flight mass separators are connected across an ion gate. Moreover, it is preferable that the mass spectrometer used here utilizes an ion source that tends to generate multivalent ions. As such an ion source, for example, an ion source by a so-called atmospheric pressure ionization method such as the above-mentioned ESI method can be cited.

In the polymer compound quantitative analysis method and data processing apparatus according to the present invention, the polymer compound to be analyzed is typically a sugar chain or a glycopeptide. This is because in sugar chains and glycopeptides, a common product ion generated by cleavage of the precursor ion is known regardless of differences in the structures of the sugar chains and glycopeptides.
For example, in a labeled N-linked sugar chain labeled with an N-linked glycopeptide, 2-aminopyridine, or the like, the mass-to-charge ratio m / w derived from the core structure (3Hex-2HexNac) of the N-linked sugar chain Product ions whose z is 138 may be set as a common product ion (see Patent Document 1). In addition, PMP-labeled O-linked sugar chains labeled with 1-phenyl-3-methyl-5-pyrazolone (PMP) are preferentially eliminated by cleavage. A product ion having a mass-to-charge ratio m / z of 175 derived from a complete PMP may be used as a common product ion.

When a plurality of multivalent ions having different ionic valences are generated from a certain polymer compound, it is possible to set MRM transitions for all the ionic valences in a predetermined range and perform MRM measurement. When the number of MRM transitions per unit is large, the time for MRM measurement under one MRM transition is shortened, or the time interval for repeating the MRM measurement under the same MRM transition is increased. The former leads to a decrease in analytical sensitivity, and the latter leads to a decrease in quantitative accuracy. Therefore, in the data processing apparatus according to the present invention, it is preferable that an analyst can input and set compound information, information on ion valence, and information on product ions. For specific polymer compounds such as sugar chains and glycopeptides, the analyst can estimate to some extent the range of ionic valences of ions generated, for example, during ionization by the ESI method based on a priori information. Therefore, by inputting and setting information based on such estimation, it is possible to avoid useless measurement in which significant data cannot be obtained.

In the polymer compound quantitative analysis method according to the present invention, in the peak information acquisition step, the peak areas derived from the target compound appearing in the mass chromatograms obtained by the plurality of MRM measurements for one target compound are totaled. At this time, the peak area obtained from the mass chromatogram obtained by one MRM measurement reflects the amount of ions having a certain ion valence derived from the target compound. Therefore, by summing multiple peak areas that reflect the amount of ions with different ionic valences derived from the target compound, a total value that reflects the amount of ions that does not depend on the ionic valence is calculated. Used for quantification of target compounds. For example, when the sample is a mixture of sugar chains or glycopeptides, measurement is performed so as to cover almost all sugar chains or glycopeptides contained in the sample, and the total value of each peak area is obtained based on the results. Thus, the abundance ratio of each sugar chain or glycopeptide in the sample can be calculated.

In order to execute such calculation and output the result, in the data processing apparatus for quantitative analysis of the polymer compound according to the present invention,
The sample is a mixture of sugar chains or glycopeptides,
The peak information acquisition unit acquires a total value of a plurality of sugar chains or glycopeptides contained in the sample,
The quantitative processing unit is based on a total value for any sugar chain or glycopeptide obtained by the peak information acquisition unit and a total value obtained by summing the total values of a plurality of sugar chains or glycopeptides contained in the sample. And determining the abundance ratio of the arbitrary sugar chain or glycopeptide,
It is good to set it as the structure further provided with the fixed_quantity | quantitative_assay result presentation part which presents the abundance ratio of arbitrary sugar_chain | carbohydrate or glycopeptide obtained by the said fixed_quantity | quantitative_assay processing part on the screen of a display part in a graph format or a table | surface form.

In addition, in order to obtain data to be processed in the polymer compound quantitative analysis method and data processing apparatus according to the present invention, as described above, a sample containing a compound separated by a liquid chromatograph is used as a mass spectrometer. Although it is not essential to use the LC-MS introduced into the LC-MS, when performing multi-component simultaneous analysis, particularly when quantifying compounds having the same mass and various structures such as sugar chains, the LC-MS Preferably used to obtain data.

According to the method for quantitative analysis of a polymer compound and a data processing apparatus for the quantitative analysis according to the present invention, a polymer compound such as a sugar chain or a glycopeptide, in which a plurality of types of polyvalent ions are easily generated upon ionization. Can be efficiently and accurately quantified using MRM measurement in a mass spectrometer capable of MS / MS analysis. In particular, the use of LC-MS enables simultaneous multi-component analysis of a sample such as a sugar chain or a mixture of glycopeptides, thereby improving the throughput of quantitative analysis of these polymer compounds and the time required for analysis. Savings and labor savings can be achieved.

BRIEF DESCRIPTION OF THE DRAWINGS The schematic block diagram of the quantitative analyzer which is one Example of this invention. Explanatory drawing which shows an example of the string | linking of the analysis object compound, precursor ion, and product ion in the quantitative analysis apparatus of a present Example. The figure which shows an example of the combination of the precursor ion which is a multivalent ion and the common product ion in the quantitative analysis apparatus of a present Example. The figure which shows an example of the mass chromatogram in the several MRM transition derived from the same sugar_chain | carbohydrate obtained in the quantitative analyzer of a present Example. The figure which shows another example of the mass chromatogram in the several MRM transition derived from the same sugar_chain | carbohydrate obtained in the quantitative analyzer of a present Example. The figure which shows an example of the graph of the abundance ratio of the sugar chain which is a quantitative analysis result in the quantitative analysis apparatus of a present Example, and a table | surface.

Hereinafter, an embodiment of a quantitative analysis method for a polymer compound according to the present invention and a data processing apparatus for carrying out the method will be described with reference to the accompanying drawings. Here, as an example, a mixture of sugar chains or glycopeptides is used as a sample, and the sugar chains or glycopeptides in the sample are quantified.
FIG. 1 is a schematic configuration diagram of a quantitative analysis apparatus of the present embodiment including a data processing apparatus according to the present invention.

This quantitative analyzer is obtained by the measuring unit 2 including the liquid chromatograph (LC) 20 and the tandem quadrupole mass spectrometer (MS / MS) 21 and the operation of the measuring unit 2 and obtained by the measuring unit 2. A control / processing unit 3 for processing the received data, and an input unit 4 and a display unit 5 which are user interfaces.

Although not shown, the liquid chromatograph 20 includes a liquid feed pump for feeding a mobile phase, an injector for injecting a sample into the mobile phase, a column for separating components in the sample in the time direction, and the like. On the other hand, the tandem quadrupole mass spectrometer 21 uses an ESI ion source, a front quadrupole mass filter, a collision cell, a rear quadrupole mass filter, to ionize components in the liquid sample eluted from the column. Including detectors. The tandem quadrupole mass spectrometer 21 may be another type of mass spectrometer capable of MS / MS analysis, such as a Q-TOF mass spectrometer.

Usually, the control / processing unit 3 is a personal computer or a higher-performance workstation (however, they are not necessarily one, and may be a plurality of computers connected to each other). Each of the functional blocks included in the control / processing unit 3 is implemented by one or more installed dedicated control / processing software operating on the computer.

Specifically, the control / processing unit 3 includes an analysis control unit 31, a data collection unit 32, and a data storage as functional blocks (reference numeral 3A in FIG. 1) embodied by existing control / processing software. A section 33, a chromatogram creation section 34, and a peak area calculation section 35. In addition, the control / processing unit 3 executes a characteristic data process, which will be described later, as a functional block (reference numeral 3B in FIG. 1) embodied by newly prepared control / processing software. Measurement target setting unit 300 including m / z calculation unit 301, measurement condition setting unit including multivalent ion information file creation unit 303, multivalent ion information file storage unit 304, method file creation unit 305, and method file storage unit 306 302, a peak area summation unit 311, a multivalent ion quantitative calculation unit 310 including a quantitative value calculation unit 312, and a quantitative output information creation unit 313. Naturally, the functional blocks indicated by reference numerals 3A and 3B in FIG. 1 may be realized by a single control / processing software.

Samples to be measured are, for example, N-linked glycopeptide mixtures, labeled N-linked sugar chain mixtures, PMP-labeled O-linked sugar chain mixtures, and the like. All of them are prepared through a predetermined pretreatment. The N-linked glycopeptide is prepared, for example, by digesting a glycoprotein such as an antibody with an appropriate enzyme such as trypsin and complying with the method described in Non-Patent Document 3. The labeled N-linked sugar chain is prepared by, for example, treating a sugar chain with glycanase with PNGase and then labeling with 2-aminopyridine or the like. The PMP-labeled O-linked sugar chain is prepared, for example, by non-reductive alkaline β elimination and labeling with PMP according to the method described in Non-Patent Document 4.

Prior to measuring the sample as described above by the measurement unit 2, the analyst inputs measurement conditions from the input unit 4 such as a keyboard. First, the analyst should examine the structure of various sugar chains that are assumed to be contained in the sample, the ionic valence of ions (precursor ions) that are assumed to be generated from the sugar chains, and the structure of the sugar chains. The mass-to-charge ratio of the product ions that appear in common regardless of, is input and set.
Specifically, when the analyst performs a predetermined operation with the input unit 4, the measurement target setting unit 300 displays a list of various sugar chain structures on the screen of the display unit 5. The analyst selects a plurality of sugar chain structures assumed to be included in the sample from the list, and inputs a plurality of ion valences for each. Since the mass is known for each glycan structure, the precursor m / z calculation unit 301 calculates the mass-to-charge ratio of ions derived from the glycan for each glycan structure from the mass and the input valence. Calculated as the precursor ion mass to charge ratio.

The measurement target setting unit 300 displays a candidate list of product ion mass-to-charge ratios on the screen of the display unit 5, and the analyst selects an appropriate mass-to-charge ratio from the list according to the type of sample. Since the mass-to-charge ratio of the product ion usually depends on the backbone structure of the sugar chain, when the component in the sample is an N-linked glycopeptide or a labeled N-linked sugar chain, it is disclosed in Patent Document 1. As shown, m / z 138 may be selected as the product ion mass-to-charge ratio. This is an ion derived from the core structure (3Hex-2HexNac) of the N-linked sugar chain. Further, when the component in the sample is a PMP-labeled O-linked sugar chain, as disclosed in Japanese Patent Application No. 2015-88346, which is a prior application by the present applicant, m as the product ion mass-to-charge ratio. / z175 may be selected. This is an ion derived from PMP, which is a complete body that is preferentially eliminated by cleavage.

Since the mass-to-charge ratio of many precursor ions and the mass-to-charge ratio of product ions are determined by the above processing, a large number of MRM transitions combining the mass-to-charge ratio of one precursor ion and the mass-to-charge ratio of one product ion are combined. Is decided. FIG. 3 is a diagram showing an example of a combination of a precursor ion that is a multivalent ion and a common product ion, that is, an MRM transition. In FIG. 3, for example, the first row shows a combination of a precursor ion having a mass-to-charge ratio of m / z 986.7225, which is a trivalent ion, and a product ion having a mass-to-charge ratio of m / z 138.

Further, the analyst inputs a measurement time range and a measurement condition such as an MRM transition of the MRM measurement executed within the measurement time range on the measurement condition setting screen displayed on the screen of the display unit 5 by the measurement condition setting unit 302. Input by part 4. Usually, the measurement time range is appropriately determined based on a known retention time of a sugar chain.

In response to the setting of the measurement conditions as described above, the method file creation unit 305 repeatedly executes MRM measurement for a predetermined one or more MRM transitions in a time range from a predetermined start time to an end time based on the sample injection. Create a method file to control each part. If only the MRM measurement for one MRM transition is set in a certain measurement time range, the MRM measurement for that one MRM transition is repeatedly executed. If MRM measurement for a plurality of MRM transitions is set in a certain measurement time range, a cycle of executing MRM measurement for each of the plurality of MRM transitions once is repeated. The created method file is stored in the method file storage unit 306.

In addition, the multivalent ion information file creation unit 303 includes a unique ID assigned to each sugar chain structure, a mass-to-charge ratio of a plurality of precursor ions having different ion valences derived from the sugar chain, and a common A multivalent ion information file that links the mass-to-charge ratio of product ions is created and stored in the multivalent ion information file storage unit 304.

FIG. 2 is an explanatory diagram showing an example of this association. In this example, the unique ID that can identify the sugar chain structure, the mass-to-charge ratio of the three types of precursor ions having ionic valences of 2, 3, and 4, and the mass of one product ion common to these precursor ions The charge ratio is tied. In this case, three types of MRM transitions exist for this sugar chain. The file for performing such association is created because the method file described above associates the measurement time range with the MRM transition, but the MRM transition and the compound (in this case, sugar chain or sugar This is because there is no correspondence with the peptide structure.

For example, when an analysis start is instructed by an analyst, the analysis control unit 31 controls the measurement unit 2 according to the measurement conditions in the method file stored in the method file storage unit 306, and performs LC / MS analysis on the sample. . In other words, the tandem quadrupole mass spectrometer 21 sequentially performs MRM measurements for various MRM transitions on the components that are sequentially eluted from the column of the liquid chromatograph 20. The data collection unit 32 collects data obtained from the tandem quadrupole mass spectrometer 21 and stores it in the data storage unit 33 as one data file.

When the analyst instructs the execution of the quantitative process after the measurement is completed, the chromatogram creation unit 34 reads the designated data file from the data storage unit 33, and converts the data obtained by the MRM measurement into each MRM transition. Based on this, a mass chromatogram showing the relationship between time and signal intensity is created.

4 and 5 show examples of mass chromatograms created in this way. In this example, an isolated peak appears in the mass chromatogram, and it is clear that this peak is derived from the target (assumed) sugar chain or glycopeptide. However, in some cases, a plurality of peaks derived from a plurality of sugar chains may appear on one mass chromatogram. This is the case, for example, when there are a plurality of sugar chains or glycopeptides having the same components but different only in structure. In such a case, it is necessary to identify and separate the target sugar chain or glycopeptide from other sugar chains or glycopeptides. Therefore, the analyst displays each mass chromatogram on the screen of the display unit 5, confirms the peak on the screen, and detects the peak of the target sugar chain or glycopeptide as necessary. Specify the allowable range of detection time.

Then, when the analyst instructs the continuation of the quantitative process, the peak area calculation unit 35 detects a peak within the allowable range of each detection time in each mass chromatogram, and calculates the peak area. In the case of normal quantitative analysis that is not multivalent ions (that is, assuming only monovalent), the quantitative value of the target compound is calculated based on this peak area value. On the other hand, in the quantitative analysis assuming multivalent ions, the following data processing is performed.

In other words, the peak area summing unit 311 in the multivalent ion quantitative calculation unit 310 reads the multivalent ion information file in which the association information is stored from the multivalent ion information file storage unit 304. Based on the association information, a plurality of MRM transitions associated with the same sugar chain or glycopeptide are recognized, and a plurality of sugar chains or glycopeptides derived from the same sugar chain or glycopeptide are identified for each same sugar chain or glycopeptide. The peak areas calculated in the mass chromatogram for the MRM transitions are summed, and the sum of the peak areas is calculated.
For example, in the example of FIG. 4, it is found that the mass chromatograms for the two MRM transitions with

valences

3 and 4 are derived from the same sugar chain based on the association information. Therefore, the peak areas obtained in the two mass chromatograms are summed up to obtain the sum of the peak areas. The same applies to the example shown in FIG.

In this way, the peak area summation unit 311 obtains the peak area summation values for all sugar chains set by the analyst before the measurement is executed. Unless there is a significant leak in the ion valence specified by the analyst, the combined peak area value should correspond to the relative content of the sugar chain or glycopeptide. Therefore, the quantitative value calculation unit 312 obtains a peak total area value obtained by summing up the peak area total values in all sugar chains or glycopeptides, and the ratio of the peak area total value in each sugar chain or glycopeptide to the peak total area value. Each (% value) is calculated as a quantitative value.

If the sugar chain or glycopeptide set by the analyst before the execution of the measurement covers the sugar chain or glycopeptide contained in the sample, the ratio of the peak area total value calculated as described above is The abundance ratio in the sample of the sugar chain or glycopeptide is shown. Therefore, the quantitative output information creation unit 313 aggregates the abundance ratios calculated for each sugar chain or glycopeptide in the form of a graph or a table, and displays this on the screen of the display unit 5 as a quantitative analysis result.

FIG. 6 is an example of a quantitative analysis result displayed on the screen of the display unit 5. In this example, the abundance ratios and standard deviations for 33 types of sugar chains are indicated by numerical values in the table below, and the abundance ratios for the structures of the 33 types of sugar chains are indicated by bar graphs above. The analyst can intuitively grasp the abundance ratio of sugar chains and glycopeptides contained in the sample by viewing this display. Of course, instead of exhaustively displaying the abundance ratio of all sugar chains and glycopeptides in this way, only the abundance ratio of specific sugar chains and glycopeptides specified by the analyst may be displayed. Good. Alternatively, only those whose abundance ratio exceeds or falls below a predetermined value may be selected and displayed.

In the description of the above examples, the present invention has been applied to the quantification of sugar chains and glycopeptides. However, the present invention is generally applied to the quantification of types of polymer compounds that can generate a common product ion regardless of the chemical structure. The invention is applicable. In addition, in the quantitative analysis apparatus of the above example, the mass analysis was performed after temporally separating the components in the sample with a liquid chromatograph, but the number of components contained in the sample was small, and the mass was analyzed. If different components having the same structure but different structures are not included, the sample can be directly introduced into the mass spectrometer without passing through the liquid chromatograph for mass analysis.

Further, the above-described embodiment is merely an example of the present invention, and it will be understood that the present invention is encompassed in the scope of the claims of the present application even if appropriate modifications, corrections, additions, etc. are made within the scope of the present invention.

2 ... Measurement unit 20 ... Liquid chromatograph 21 ... Tandem quadrupole mass spectrometer 3 ... Control / processing unit 31 ... Analysis control unit 32 ... Data collection unit 33 ... Data storage unit 34 ... Chromatogram creation unit 35 ... Peak area Calculation unit 300 ... Measurement object setting unit 301 ... Precursor m / z calculation unit 302 ... Measurement condition setting unit 303 ... Multivalent ion information file creation unit 304 ... Multivalent ion information file storage unit 305 ... Method file creation unit 306 ... Method file Storage unit 310 ... multivalent ion quantitative calculation unit 311 ... peak area summation unit 312 ... quantitative value calculation unit 313 ... quantitative output information creation unit 4 ... input unit 5 ... display unit

Claims

A quantitative analysis method for quantifying a polymer compound in a sample using a mass spectrometer capable of MS / MS analysis,
a) With respect to a polymer compound to be analyzed or assumed to be analyzed, a plurality of precursor ions having different ionic valences derived from the compound and a plurality of MRM measurements targeting one common product ion are used. A peak information acquisition step for calculating a total value of the peak areas appearing in the obtained plurality of mass chromatograms,
b) Using the total value obtained in the peak information acquisition step, a quantitative step for performing a quantitative calculation of a polymer compound that is an analysis target or assumed as an analysis target;
A method for quantitative analysis of a polymer compound, comprising:
A method for quantitative analysis of a polymer compound according to claim 1,
The method for quantitative analysis of a polymer compound, wherein the polymer compound is a sugar chain or a glycopeptide.
A method for quantitative analysis of a polymer compound according to claim 2,
The sample is a mixture of sugar chains or glycopeptides,
In the peak information acquisition step, a total value of a plurality of sugar chains or glycopeptides contained in the sample is acquired,
In the quantification step, based on the total value for any sugar chain or glycopeptide obtained in the peak information acquisition step, and the total value obtained by summing the total values of a plurality of sugar chains or glycopeptides contained in the sample The abundance ratio of the arbitrary sugar chain or glycopeptide is determined,
Quantification of a polymer compound, further comprising a quantification result presentation step of presenting an abundance ratio of any sugar chain or glycopeptide obtained in the quantification step in a graph format or a table format on a screen of a display unit Analysis method.
A method for quantitative analysis of a polymer compound according to any one of claims 1 to 3,
A method for quantitative analysis of a polymer compound, comprising: measuring a polymer compound in a sample using a liquid chromatograph mass spectrometer having a liquid chromatograph connected to a front stage of the mass spectrometer.
A data processing apparatus for quantifying a polymer compound in a sample based on data obtained using a mass spectrometer capable of MS / MS analysis,
a) With respect to a polymer compound to be analyzed or assumed to be analyzed, a plurality of precursor ions having different ionic valences derived from the compound and a plurality of MRM measurements targeting one common product ion are used. A peak information acquisition unit for calculating a total value of the peak areas appearing in the obtained plurality of mass chromatograms;
b) Using the total value obtained by the peak information acquisition unit, a quantitative processing unit that performs a quantitative calculation of a polymer compound that is an analysis target or is assumed to be an analysis target;
A data processing apparatus comprising:
The data processing apparatus according to claim 5, wherein
The data processing apparatus, wherein the polymer compound is a sugar chain or a glycopeptide.
The data processing apparatus according to claim 6, wherein
The sample is a mixture of sugar chains or glycopeptides,
The peak information acquisition unit acquires a total value of a plurality of sugar chains or glycopeptides contained in the sample,
The quantitative processing unit is based on a total value for any sugar chain or glycopeptide obtained by the peak information acquisition unit and a total value obtained by summing the total values of a plurality of sugar chains or glycopeptides contained in the sample. And determining the abundance ratio of the arbitrary sugar chain or glycopeptide,
A data processing apparatus, further comprising: a quantitative result presenting unit that presents an abundance ratio of any sugar chain or glycopeptide obtained in the quantitative processing unit on a screen of a display unit in a graph format or a table format .